Objective To establish a RP-HPLC method for the determination of drug release in vitro of atractylenolide Ⅰ liposomes. Methods The release behavior of the drug from liposomes was studied by the third method for dissolution. ZORBAX C18 column (4.6 mm × 250 mm, 5 μm) was used with a mobile phase of Methanol-Acetonitrile-0.2% from liposomes in vitro fitted the log-normal distribution equation and had a property of sustained release. Conclusion The method is simple, fast and selective. It is suitable for the determination of release profile in vitro of atractylenolide Ⅰ liposomes.

AIM:To investigate molecular regulatory mechanism of K-ras to girdin protein in COS7 cells and expression of K-ras and girdin in colorectal carcinoma tissues .METHODS:The lentiviral vector carrying K-ras gene was constructed and transfected in the COS 7 cells.The expression of K-ras, girdin proteins and other related proteins in COS 7 cells and colorectal carcinoma tissues was observed by Western blot .RESULTS:The COS7 cells with K-ras over-expres-sion showed an irregular cell morphology .The results of Western blot indicated that the downstream signal protein levels of p-ERK1/2, p-Stat3 and girdin were significantly increased in the COS 7 cells with K-ras over-expression.Transfection with the K-ras siRNA into the COS7 cells significantly reduced the protein levels of p-ERK1/2, p-Stat3 and girdin.In the color-ectal carcinoma tissues (7 cases), 5 cases had higher expression of K-ras and girdin compared with pericarcinous tissues . CONCLUSION:K-ras regulates girdin expression through the signal pathway of K-ras-ERK1/2-Stat3-girdin.

Lactate and succinate were produced by Corynebacterium acetoacidophilum from glucose under oxygen deprivation conditions. To construct knockout mutant, lactate dehydrogenase gene (ldh) of C. acetoacidophilum was deleted by double-crossover chromosome replacement with sacB gene. Comparing with the wild strain ATCC13870, ldhA-deficent mutant produced no lactate with glucose consumption rate decreased by 29.3%, while succinate and acetate concentrations were increased by 45.6% and 182%, respectively. Moreover, the NADH/NAD+ rate was less than 1 (about 0.7), and the activities of phosphoenolpyruvate carboxylase and acetate kinase of the ldhA-deficent mutant were enhanced by 84% and 12 times, respectively. Our studies show that succinicate and acetate production pathways are strengthened by blocking lactate synthesis. It also suggests that improving NADH supply and eliminating acetate generation are alternative strategies to get high succinate-producer.
Corynebacterium glutamicum ; genetics ; metabolism ; Glucose ; Industrial Microbiology ; L-Lactate Dehydrogenase ; genetics ; metabolism ; Lactic Acid ; metabolism ; Oxygen ; metabolism ; Phosphoenolpyruvate Carboxylase ; Succinic Acid ; metabolism

In anaerobic bottles fermentation,glucose,fructose,xylose,lactose,maltose,sucrose and sugar alcohols could be used to produce succinic acid with Actinobacillus succinogenes. When sorbitol was utilized as the carbon source in the batch fermentation,more succinate and ethanol were produced compared with those using glucose,while producing less acetate and formate. The metabolic flux analysis results showed that the flux partitioning at PEP node was stable when glucose was replaced by sorbitol,but the flux partitioning at PYR and AcCoA nodes changed a lot because more reducing power(NADH) was generated to meet the more requirement the synthesis of succinate and ethanol.

Objective To optimize the prescription and preparation technclogy of ligustilide liposomes. Methods Ligustilide liposomes were prepared by ethanol injection method. The encapsulation efficiency was taken as inspection target and the preparation of liposomes was optimized by orthogonal design. HPLC was used to measure the encapsulation efficiency. Results The best prescription was ligustilide-lecithin (1:10),lecithin-cholesterol (2:1),the water phosphate buffer solution (pH 7.5),the hydration temperature was 35 ℃. Conclusion The optimized formulation of ligustilide is reasonable in prescription and practicable in technology.

Objective To observe the biological role and the underlying mechanisms of miR-132 in liver cancer on invasion and metastasis.Methods Real-time quantitative polymerase chain reaction (RT-qPCR) analysis was used to examine the expression of miR-132 in four liver cancer cell lines (MHCC97H,HCCLYH,MHCC97L and SMMC-7721),a normal liver cell line HL-7702,and in liver tumor tissues with or without metastases.The biological effects of miR-132 transfection on human liver can-cer cells were assessed by wound assay,matrigel counting and in vivo experiments in nude mice.Western blotting was used to detect the expression of E-cadherin,α-cadherin,vimentin,fibronectin and ZEB2 in li-ver cancer cells.Immunohistochemistry was used to detect positive expression of ZEB2 in xenograft tumors.Results The expressions of miR-132 were downregulated in the four liver cancer cell lines when compared with the normal liver cell line (P ＜ 0.05),and in the liver cancer tissues with distant metastases when compared with the tissues without metastases (P ＜ 0.05).After transfection,ectopic expressions of miR-132 markedly inhibited cell migration and invasion in liver cancer cells.When compared with the control group,the expressions of E-cadherin and α-cadherin in the miR-132 transfection group were significantly increased,but the expressions of vimentin,fibronectin and ZEB2 were decreased.In addition,the numbers of metastatic lung lesions in nude mice in the miR-132 transfection group was markedly decreased when compared with the control group.The expressions of ZEB2 in the miR-132 transfection group was also significantly decreased when compared with the control group.Conclusions Transfection of miR-132 effectively inhibited invasion and metastasis of liver cancer cells in vitro and in vivo.miR-132 may become a new target for regulation of gene expression in liver cancer.

Objective To establish a HPLC method for simultaneous determination of chlorogenic acid and luteoloside in the leaves of“Wudang No.II” flos lonicerae. Methods Phenomenex C18(4. 6 mmí250 mm, 5μm) was used;the mobile phase was acetonitrile( A) and 0. 4% phosphoric acid aqueous solution( B) by gradient elution mode; the detection wavelength was 350 nm and the flow rate was 0. 8 mL·min-1;the column temperature was set at 32℃. Results The calibration curve of chlorogenic acid and luteoloside was linear in the range of 0. 285-2. 280μg(r=0. 999 3), and 0. 124-1. 240μg(r=0. 999 4), respectively. The mean recovery of chlorogenic acid and luteoloside was 98. 9%, RSD=1. 59% and 98. 8%, RSD=1. 84%, respectively. Conclusion This method was found to be accurate, quick and reproducible. It can be used for simultaneous determination of chlorogenic acid and luteoloside in the leaves of “Wudang NO.II”flos lonicerae.

Objective To study the effect of macroporous adsorptive resins on the decoloration technology ofLonicera japonica Thunb. polysaccharides(FLP).Methods The effects of 6 kinds of macroporous adsorptive resins i.e. HPD-400A, AB-8, HPD-750, HPD-100, D3520, D301T, S8 on the decolorization technology ofLonicera japonica Thunb. polysaccharides were compared with single factor test in terms of temperature, polysaccharide concentration, pH, adsorption flow, and eluant.Results The decolorization rate and polysaccharide retention rate of S8 were optimal. The best decoloration conditions were as follows: temperature of 40℃, polysaccharide concentration of 5 mg/ml, pH value of 6, flow rate of 1 ml/min, distilled water with pH=6 as eluant. The adsorption rate was 83.2%,and polysaccharide retention rate was 72.1%.Conclusion High decolorization ratio and the high retention rate ofLonicera japonica Thunb. could be obtained by means of decoloration with S8 macroporous adsorptive resins..

Objective To analyze the epidemiology and clinical features of the critical congenital heart diseases in neonates,and to summarize the main points on pre-operative treatments.Methods We retrospectively analyzed all critical congenital heart disease in newborns admitted to CICU from June 2014to June 2015.Depict entity distribution,the main symptoms,and the key points on their treatments,also the indi-cation of intubation and their operation time were summarized.Results In totally 96cases,transposition of the great artery,with and without the intact ventricle septum,was the biggest category and counts for 49%in our group.Severe cyanosis was the main symptom for 62.5% of all cases.Another key symptom was the heart failure(33.3%).Eight-seven cases were intubated during their treatments,in which 41were intubated as soon as they were admitted and 40cases were done in the first 24hours after their admission.One case died before treatment due to interrupted aortic arch.All the rest received operations during their hospital stay. Conclusion Transposition of great artery is the dominating entity in critical congenital heart diseases in new-borns;severe cyanosis is the main symptom.Treatment should be based on each characteristic anatomy and hemodynamics features.Rigorous and dynamic monitoring on the homeostasis and metabolic index determines the indication of intubation and surgery time.

Acute appendicitis is one of the most common acute abdominal diseases in surgery. Due to its urgent onset and rapid progress, timely diagnosis and early treatment are beneficial to the prognosis of patients. The diagnosis of acute appendicitis should be based on the characteristics of different populations, based on clinical symptoms, physical examination, laboratory examination, and imaging methods. Anti-infection treatment should be started as soon as possible in the early stage, and antibiotics should choose effective drug treatment with both anaerobic bacteria and aerobic bacterial infection. Appendectomy is still the most effective treatment for acute appendicitis. In the case of uncomplicated abdominal infection such as simple and non-perforated appendicitis, antibiotic treatment may be chosen, but the development needs to be closely observed. Laparoscopic appendectomy is the first choice, but in pregnancy patients should be cautious. Emergency surgery is required for acute perforated appendicitis and attention should be paid to intraoperative abdominal flushing. Immediate surgical treatment of appendiceal abscess may involve the risk of partial ileectomy. Antibiotic treatment is recommended, with percutaneous abscess puncture and drainage if necessary.